title: In vitro toxicity of melamine against Tetrahymena pyriformis cells.
abstract: This study assessed the toxicity of melamine against the unicellular eukaryotic system of Tetrahymena (T.) pyriformis exposed to 0, 0.05, 0.25, 0.5, 2.5, and 5 mg/mL of melamine. Cell growth curves of different cultures, the half maximum inhibition concentration (IC(50)) value of melamine, and morphological changes in cells were obtained via optical and transmission electron microscopic observation. The effects of eleven melamine concentrations, 0.25, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5 and 5 mg/mL, on protein expression levels of T. pyriformis were examined using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The results showed an obvious inhibitory effect of melamine on the growth of eukaryotic cells. Cell growth dynamics indicated that the IC(50) value of melamine on T. pyriformis was 0.82 mg/mL. The cellular morphology was also affected in a concentration-dependent manner, with characteristics of atrophy or cell damage developing in the presence of melamine. The relative contents of the top four main proteins corresponding to peak mass-to-charge ratios (m/z) of 4466, m/z 6455, m/z 6514, and m/z 7772 in the MALDI-TOF-MS spectra were all found to be closely correlated with the melamine concentrations. In conclusion, exposure of eukaryotic cells to melamine could inhibit cell growth, cause changes in cytomorphology and even disturb the expression of proteins in a concentration-dependent manner. The described method of examining four sensitive proteins affected by melamine was also proposed to be used in a preliminary study to identify protein biomarkers in T. pyriformis.
authors.affiliation: College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.
authors.forename: Zhengfang
authors.initials: Z
authors.lastname: Wang
authors.forename: Liben
authors.initials: L
authors.lastname: Chen
authors.forename: Rashad
authors.initials: R
authors.lastname: Al-Kasir
authors.forename: Bo
authors.initials: B
authors.lastname: Han
mesh_terms: D000821:Animal Feed / Q000032:analysis / Q000633:toxicity; D015415:Biomarkers / Q000032:analysis; D005503:Food Additives / Q000032:analysis / Q000633:toxicity; D020128:Inhibitory Concentration 50; D046529:Microscopy, Electron, Transmission; D015800:Protozoan Proteins / Q000032:analysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013769:Tetrahymena pyriformis / Q000166:cytology / Q000187:drug effects*; D014227:Triazines / Q000633:toxicity*
chemical_list: D015415:Biomarkers; D005503:Food Additives; D015800:Protozoan Proteins; D014227:Triazines; C011907:melamine